6alpha, 16alpha-dimethyl-11-oxygenated-17alpha-hydroxy-unsaturated-pregnane-3, 20-diones



3,005,838 60,16a DlMETI-IYL 11 QXYGENATED 17cc- RQXY UNSATURATEDPREGNANE- 3,20-DIONES Frank H. Lincoln, Kalamazoo, William P. Schneider,Kalamazoo Township, Kalamazoo County, and George B. Spero, Kalamazoo,Mich, assignors to The Upjohn Company, Kalamazoo, Mich, a corporation ofMichigan No Drawing Filed Sept. 15, 1958, Ser. No. 760,849 1 Claim. (Cl.260-39145) wherein X is a hydrogen, chlorine or fluorine atom, R is analiphatic or aryl radical, preferably hydrocarbon containing from 1 to12 carbon atoms, inclusive, e.g., tolyl or methyl, and the dotted linerepresents a A -double bond 3,005,838 Patented Get. 24, 1961 which mayor may not be present, i.e., the formulae represent both the A and the Acompounds.

The novel compounds of this invention represented by Formula IV and thecorresponding ll-keto compounds (V) are highly active anti-inflammatoryagents with improved ratio of therapeutic activity to undesirable sideeffects, e.g., gastro-intestinal disturbances, salt retention, edema,etc., known to exist with similar known physio logically activesteroids. These compounds are useful in the treatment of variousinflammatory conditions of the skin, eyes, respiratory tract and thebones and internal organs of the animal organism due to bacterial orviral infections, contact dermatitis and allergic reactions. For thispurpose, they may be administered in conventional dosage forms, e.g.,pills, tablets, capsules, syrups or eliXirs for oral use, liquid formswhich are adaptable for injectable products, or topically in the form ofointments, creams, sprays, lotions and the like, with or withoutcoacting antibiotics, e.g., the penicillins, neomycin, tetracyclin,chloromycetin and novobiocin, or other materials forming advantageouscombinations therewith.

The novel compounds of this invention (IV) are prepared from6or,16or-dirnethyl-1 1B,17or,21-trihydroXy-4-pregnone-3,20-dione,6a,16a-dimethyl-1 1fi,17a,21-trihydroxy- 1,4-pregnadiene-3,ZO-dione andthe corresponding 9afiuoro and 9a-chloro compounds by esterifying the 21hydroxy group of these compounds with a sulfonating agent, e.g., analkyl or aryl, preferably hydrocarbon containing from 1 to 12 carbonatoms, inclusive, sulfonyl chloride or bromide, to produce thecorresponding 21- sulfonate ester (II). The usual reaction conditionsare employed, e.g., the steroid is dissolved in pyridine or an inertorganic solvent containing pyridine or like amine and reacted with thehydrogen halide formed in the reaction, at preferably room temperatureor below. The isolated 21-sulfonate ester is then reacted with analkalimetal iodide to produce the corresponding 21-iodo compound (IH).The 21-i0dine atom is then removed from the steroid molecule bytreatment with one of the usual reducing agents, e.g., sodiumthiosulfate or other reducing agents known to remove an iodine atom,thus producing 6a,16o-dimethyl11,8,17a-dihydroxy-4-pregnene-3,20 dione,6m,16a-dimethyl-1 1,8,17a-dihydroXy-1,4-pregnadione-3,20-dione, thecorresponding 9a-fluoro and 9zx-Chi01'0 compounds, respectively, (IV)depending upon the starting 2l-hydroxy compound (I). The ll-ketocompounds corresponding to each of the above compounds are prepared byoxidizing the llp-hydroxy group with, e.g., chromic acid, N-haloamide orN-haloimide in pyridine or like amine.

The starting 6a,16u-dimethyl-l1fi,17a,21-trihydroXy-4- pregnene3,20-dione, 6u,16x-dimethyl-11/3,l7u,21-trihydroXy-l,4-pregnadiene-3,ZO-dione, thecorresponding 9afluoro compounds and the corresponding 9u-chlorocompounds are prepared from the known 16u-methylprogesterone (l) [Markeret al., J. Am. Chem. Soc., 64, 1280 (1942)], as shown in thepreparations hereinafter, e.g., bioconversion of 16a-methylprogesteronewith Rhizopus arrhizus according to the method of US. 2,602,769 toproduce 1lu-hydrony-l6u-methylprogesterone; esterification of thella-hydroxy group with acetic anhydride in pyridine to produce11a-hydroxy-l6u-methylprogesterone 11-acetate; ketalization of the 3 and20 keto groups of either of these compounds with ethylene glycolaccording to methods well known in the art to produce the corresponding3,20-bis-(ethylene ketal). The thus obtained diketal is then reactedwith a peracid, e.g., peracetic, to produce the 50,6a-0Xid6, i.e.,5m,6oc 3p0Xy-1lot-hYdl'OXY-lGotmethylallopregnane-3,ZO-dione 3,20-bis-(ethylene ketal) or ll-acetate; reacting one of these compoundswith methyl lithium or methyl magnesium bromide to produce5a,l1a-dihydroxy 6p,16a-methylallopregnane-3,ZO-dione 3,20-bis-(ethyleneketal). The ketal group is then hydrolyzed with acid to giveSaJla-dihydroxy-6fl,16a-dimethylallopregnane-3,20-dione. In the nextstep the 11- hydroxy group is oxidized, e.g., with chromic acid, sodiumdichromate or an N-haloamide or N-haloirnide in pyridine or like amineto give a-hydroxy-6e,l6ot-dimethylallopregnene3,1l,20-trione. Thiscompound is converted with aqueous acid or base to60,16oL-diIIl6thYl-ll-k6t0- progesterone.

6oz,16u-dimethyl-1l-ketoprogesterone is converted to3,11-diketo-6a,16a-dimethyl-4,7(20)-pregnadien 21 oic acid methyl esteraccording to the procedure of US. 2,790,814, i.e., reaction with over 2molar equivalents each of sodium methoxide and methyl oxalate followedby acidification with acetic acid and then bromination with about 3molar equivalents of bromine, followed by reaction with sodium methoxideto produce 2-bromo-3,1ldiketo-Ga,16x-dimethyl-4,17(20)-pregnadien-21 oicacid methyl ester, whose 2-bromo group is then removed with zinc andacetic acid.

Conversion of 3,1l-diketo-6a,16u-dimethyl-4,17(20') pregnadien-21-oicacid methyl ester to 6a,16a-dimethyl-11fl,21-dihydroxy-4,17(20)-pregnadien-3-one is accomplished in themanner described in US. 2,844,604, i.e., the 3-keto group is protectedby conversion with pyrrolidine to a 3-enamine group and then reducingthe ll-keto and 2l-carbonyl groups with lithium aluminum hydride toproduce the S-pyrrolidyl enamine of 6a,l6vt-dirnethyl-1113,21-dihydroxy-4,17(20)-pregnadien-3-one. The enamine group is thenremoved with aqueous base to produce 6a,l6z-dimethyl-l1fl,21-dihydroxy-4,17(20) pregnadien-S-one which is thenesterified in the 2l-position with acetic anhydride and pyridine underthe usual esterification conditions to produce the corresponding21-acetate. This latter compound is then oxidatively hydroxylated withasmium tetroxide and hydrogen peroxide, an amine oxide peroxide or aryliodo oxide [U.S. 2,769,823; 2,769,825; Hogg et al., J. Am. Chem. Soc.,77, 4436 (1955)] to produce6a,l6u-dimethyl-11B,17,21-trihydroxy-4-pregnene 3,20 dione 21-acetate.The corresponding A compound is prepared by bioconverting 60,160cdimethyl 1l,8,21-dihydroxy-4,l7 -(20)-pregnadien-3-one with Septomyxaafifinis to produce 11,8,21-dihydroxy-1,4,17(20)-pregnatrien-3-one whichis then esterified in the 2l-position and the l7(20)-double bondoxidatively hydroxylated as described above to produce6m,l6a-dimethyl-l1,8,17oc,2l-trihydroxy-1,4-pregnadiene-3,20-dione21-acetate. These two compounds are hydrolyzed with sodium bicarbonateunder oxygen free conditions to produce the corresponding 21-hydroxycompounds (I, X=H).

The corresponding 9a-fiuoro and 9oc-Chl010 compounds (I, X=Cl, F), areprepared by reacting6a,16ct-dimethyll15,17u,21-trihydroxy-4-pregnene-3,ZO-dione 21 acetateand 6a,l6a-dimethyl-l1fi,l7a,2l-trihydroxy-1,4-pregnadiene-3,2()-dione2l-acetate with, for example, an N-haloamide or N-haloimide and sulfurdioxide in pyridine, ptoluene-sulfonic acid or thionyl chloride, toproduce the corresponding 9(11)-dehydro compounds. These compounds arethen reacted with hypochlorous acid to produce6a,l6ot-dimethyl-9a-chloro-l 1B, l7a,2l-trihydroxy-4-pregnene-3,20-dione 21-acetate and 6a,16 x-dimethyl-9otchloro-ll5,l7a,21-trihydroxy-1,4-pregnadiene-3,20 dione 21-acetate,respectively. These compounds or the corresponding 9oc-b1OII1Ocompounds, prepared by substituting hypobromous acid for thehypochlorous acid above, are then reacted with a base, e.g., potassiumacetate, to give the corresponding 9(11)-epoxy compounds which areconverted with anhydrous or aqueous hydrogen fluoride, according tomethods well known in the art, to 60:, 16a-dim6thYl-9a-fil1OI0-l1B,17a,21-trihydroxy-4-pregnene- 3,20-dione Zl-acetate and6a,16a-dimethyl-9a-fluoro-11 ,8,17a,21-trihydroxy-1,4-pregnadiene-3,20-dione 21-acetate, respectively.Each of these 9oz-halo-21-acetate compounds s then hydrolyzed with base,e.g., aqueous sodium bicar- PREPARATION 1 11a-hydroxy-l6ormethylpr0gester0ne To 16.6 1. of a fermentation mediumconsisting of 1.2% corn steep solids and 2% Cerelose glucose andadjusted to a pH of 4.8 to 5.0 was added an inoculum of Rhizopusnigricans (A.T.C.C. 6227b) and the medium incubated for 24 hours at atemperature of 28 C. with a rate of aeration of 5% air 'by volume perminute. To this medium was added 5 gm. of 16a-methylprogesterone [Markeret al., J. Am. Chem. Soc., 64, 1280 (1942)] dissolved in 35 ml. ofacetone. After an additional 24 hours of incubation under the sameconditions, the mycelium was filtered, washed twice, each time with :avolume of acetone approximately equal to the volume of the mycelium andthen twice with similar portions of methylene chloride. The combinedextracts were added to the beer filtrate and the whole was extractedsuccessively with 2 one-half by volume portions of methylene chlorideand then with 2 one-fourth by volume portions of methylene chloride.These extracts were washed with 2 one-tenth by volume portions of 2%aqueous sodium bicarbonate and then with 2 one-tenth by volume portionsof water. The methylene chloride extracts were then dried and thesolvent removed by distillation. The residue Was dissolved in 250 ml. ofmethylene chloride and chromatographed over a 500 gm. column ofmagnesium silicate (Florisil). The column was developed with 5 l. ofmethylene chloride, 5 l. of hexanes (Skellysolve B) plus 5% acetone, 10l. of hexanes plus 10% acetone, 51. of hexanes plus 25% acetone and 2 l.of acetone. The last 7 l. of hexanes plus 10% acetone, the hexanes plus25% acetone and the first acetone eluates were combined, freed ofsolvent, redissolved in methylene chloride and rechromatographed over400 gm. of magnesium silicate. The column was developed with 8 l. ofhexanes plus 10% acetone, 8 l. of hexanes plus 15% acetone, 4 1. ofhexanes plus 20% acetone, 4 l. of hexanes plus 25% acetone and 2.4 ml.of acetone, in that order. The combined residues from the last 4 l. ofhexanes plus 10% acetone and the first 1.6 ml. of hexanes plus 15%acetone eluates were recrystallized twice from ethyl acetate to giveIla-hydroxy-loot-methylprogesterone melting at 161 to 163 (3., having an[0:1 of plus 149 (chloroform) and the analysis below.

Analysis.-Calculated for H H O C, 76.70; H, 9.36. Found: C, 74.46; H,9.63

PREPARATION 2 11 a-hydroxy-l 605-171 ethylprogesterone 3,20-bisethyleneketrrl) A solution of 10 gm. of 1lot-hydroxy-l6ot-methylprogesterone, 1gm. of para-toluenesulfonic acid in 1,000 ml. of benzene and ml. ofethylene glycol was refluxed for 6 hours using a water trap to removethe water formed in the reaction. The solution was cooled, water wasadded and the aqueous layer was separated and washed with ether and theether extracts added to the organic layer. The combined organic layerswere successively washed with 5% sodium bicarbonate solution, saturatedsodium chloride solution, water and then dried over sodium sulfate. Thesolvents were removed by distillation and the thus-obtained residue wasrecrystallized from methanol to give l1a-hydroxy-l6a methylprogesterone3,20-bis-(ethylene diketal).

Esterification according to the method of Preparation 13 gave11a-hydroxy-16u-methylprogesterone 3,20-bis- (ethylene diketal)ll-acetate.

PREPARATION 3 11 a-hydroxy-I6u-methylprogesterone 3,20-bis-(ethyleneketal) 11 -acetate In thesame manner as shown in Preparation 2, 6 gm. of1la-hydroxy-l6a-methylprogesterone ll-acetate, dissolved in 110 ml. ofbenzene and 11 ml. of ethylene glycol, was heated to reflux in thepresence of 270 mg. of para-toluenesulfonic acid for a period of 18hours to give lloc-hYdrOXY-lGoa methylprogesterone 3,20-bis-(ethyleneketal) ll-acetate.

PREPARATION 4 5 a,6a-epoxy-1 1 a-hydroxy-I 6-methylall0pregnane-3,20-dione 3,20-bis-( ethylene ketal) 1 l-acetate PREPARATION 5 5 a,6a-@pxy-11 a-hydroxy-16u-methylallopregnane-3,20- dione 3,20-bis-(ethylene ketal)In the same manner as shown in Preparation 4, reacting11a-hydroxy-l6a-methylprogesterone 3,20-bis-(ethylene ketal) withperacetic acid and anhydrous sodium acetate in chloroform solutionproduced Sonja-epoxy- 1lu-hydroxy-16a methylpregnane-3,20 dione 3,20bis- (ethylene ketal).

PREPARATION 6 0a,] 1 a-dihydroxy-6 ,8,1 6 a-dimethylallopregnane-3,20-dione 3,20-bis-(ethylene ketal) To 5 ml. of 3 molar ether solution ofmethyl magnesium bromide was added dropwise a solution of 476 mg. of5u,6a-epoxy-11a-hydroxy-16a-methylallopregnane- 3,20-dione3,20-bis-(ethylene ketal) ll-acetate in 20 ml. of distilledtetrahydrofuran. The reaction mixture was stirred and refluxed for 17hours and was then cooled and 25 ml. of iced saturated ammonium chloridesolution was added. After stirring for a few minutes the mixture wasextracted with ether and the ether was washed with water, drived overanhydrous sodium sulfate and evaporated to dryness. The residue wascrystallized from a mixture of acetone and hexanes (Skellysolve B) togive 5a,l1a-di hydroxy-6;3,16a-dimethylallopregnane-3,20-dione 3,20-bis-(ethylene ketal).

PREPARATION 7 5 ,1 1 a-dihydr0xy-6 3,1 6 a-dimethy lallopregnane- 3,ZO-dione A solution of 459 mg. of5a,11a-dihydroxy-6,3,16adimethylallopregnane-3,20-dione3,20-bis-(ethylene ketal) in 10 ml. of acetone and 1 ml. of l N sulfuricacid was gently boiled on the steam bath for 10 minutes. The solutionwas cooled to room temperature, diluted with 20 ml. of water and thenrefrigerated. There was obtained a crystalline precipitate of5a,11a-dihydroxy- 65,16u-dimethylallopregnane3,20-dione.

PREPARATION 8 5 a-hydroxy-6 5,1 6 Ct-d imethy lall0pregnane-3,1 1,ZO-trione To a solution of 5 gm. of sodium dichromate dihydrate in ml.of glacial acetic acid was added 10 gm. of5e,1loc-dihydroxy-6p,16a-dimethylallopregnane- 3,20-dione in ml. ofacetic acid and the mixture was stirred for 1 hour at room temperature.The solution was then cooled to between 0 and 5 C. and 1.5 l. of ice andwater was added with vigorous stirring. When addition was complete, 150mg. of sodium sulfite was added and the solution was then maintained atbetween 0 and 5 C. overnight. The precipitated steroid was filtered andwashed with water and then dried. There was thus obtained substantiallypure 5a-hydroxy-6,B,l6a-dimethylallopregnane-ll1,20-trione which can befurther purified by recrystallization from methylene chloride andmethanol, after decolorization with'charcoal.

PREPARATION 9 6a,16a-dimethyl-1 1 -ket0pr0gester0ne To a stirredsolution of 4.576 gm. of Sa-hydroxy-6e,16u-dimethylallopregnane-3,11,20-trione in 440 ml. of methanol in anitrogen atmosphere was added a solution of 40 ml. of 0.1 N sodiumhydroxide, similarly freed of air with nitrogen. The mixture was stirredunder nitrogen for 16 hours and then made slightly acidic with glacialacetic acid. The solution was concentrated by distillation at reducedpressure to about 35 ml. There was thus obtained a precipitate of6a,l6oc-dimethyl-11- ketoprogesterone which was filtered, washed withcold methanol and dried.

The corresponding 6fl-isomer, i.e., 6B,16u-dimethyl-11-ketoprogesterone, is prepared by dehydrating with a mixture ofN-bromoacetamide and sulfur dioxide in pyridine instead of sodiumhydroxide. This compound is converted, according to the procedure ofPreparation 10 to 3,1 1-diketo-6,8,16a-dimethyl 4,17 (20)-pregnadien-21-oic acid methyl ester which, in turn, is converted to6u,16adimethyl-l 15,2 1-dihydroxy-4, 17( 20) -pregnadien-3 -one.

PREPARATION l0 6 a,] 6 a-dimethyl-i ,1 1 -diket0-4,1 7 (20-pregnadien-21 -0ic acid methyl ester A solution of 7.12 gm. (0.02 mole)of 60:,16oc-d-imethyl-1l-ketoprogesterone in 70 ml. of tertiary butylalcohol was prepared by heating to 55-60 with stirring under a nitrogenatmosphere. While the temperature was held at this point 11.7 gm. ofethyl oxalate was added. Thereafter 2.7 gm. of sodium methoxide inmethanol (commercial 25% solution) was added. Almost immediately theyellow precipitate of the sodium dienolate of6a,16u-dimethyl-2,21-diethoxyoxalyl-ll-ketoprogesterone began toprecipitate. The mixture was allowed to cool slowly to approximately 35C. while stirring for 15 minutes.

A solution of 2.44 gm. of anhydrous sodium acetate and 3.00 gm. ofglacial acetic acid in ml. of methanol which had been previously cooledto 10 C. was then added and the mixture stirred until the solution wasachieved. The yellow solution was cooled to 0 and to this vigorouslystirred solution was added dropwise a precooled (0 C.) solution of 9.6gm. of bromine in 96 ml. of methanol. Approximately 75 ml. of thebromine solution was added at a constant rate during 10 minutes. Therate of addition was then decreased and the remaining 21 ml. addedduring the following 10 minutes. After an additional ten-minute stirringperiod, the bromine color had essentially disappeared.

With continuous stirring and cooling a solution of 5.57 gm. of sodiummethoxide in methanol (commercial 25% solution) was added rapidly. Abright orange color developed which soon faded to yellow amber. Thecooling bath was removed, the temperature raised to 2530 C. and thesolution stirred for 1.5 hours.

16 ml. of acetic acid and 3.2 gm. of zinc dust was added to thismaterial and stirring was continued for 30 minutes. The excess zinc dustwas removed by filtration and washed with 15 to 20 ml. of freshmethanol. The combined filtrates were concentrated at reduced pressurein a 60 water bath to approximately 200 ml. The concentrate was pouredslowly with stirring into 750 ml. of ice and water. The mixture wasrefrigerated for 15 minutes and then filtered. The filter cake waswashed with 100 ml. of cold water and dried at room temperature to give3,1l-diketo-6oa,16a-dimethyl-4,17(20)-pregnadien- 21-oic acid methylester.

A 1.5 gm. portion of the crude material was dissolved in 50 ml. ofbenzene and poured onto a 75 gm. magnesium silicate (Florisil)chromatographic column. The column was eluted with 750 ml. of hexanes(Skellysolve B) plus 2% acetone, 1200 ml. of hexanes plus 5% acetone,300 ml. of hexanes plus acetone, and 150 ml. of 100% acetone. The eluatewas collected in 150 ml. fractions and evaporated. The hexanes plus 5%acetone eluted 3,11 diketo-6a,16a-dimethyl-4,l7(20)-pregnadien-2l-oicacid methyl ester which was freed of solvent by evaporation.

PREPARATION 11 6a,]6a-dimethyl-115J1-dihydr0xy-4,1 7(20) -pregnadien3-0ne A solution of 0.5 gm. of 3,11-diketo-6a,16ot-dimethyl-4,17(20)-pregnadien-2l-oic acid methyl ester, 0.5 ml. of pyrrolidine, 40ml. of benzene and 20 mg. of p-toluenesulfonic acid was heated underreflux for 1 hour. The solvent was distilled under vacuum. Triturationof the residue with methanol gave, as a yellow solid, the 3- pyrrolidylenamine of 3,11-diketo-6a,l6adimethyl-4,l7 (20)-pregnadien-21-oic acidmethyl ester.

To a suspension of 1.1 gm. of lithium aluminum hydride in 75 ml. ofanhydrous ether was added 1.3 gm. of the crude 3-pyrrolidyl enamine of3,1i-diketO-6a,16adimethyl-4,17(20)-pregnadien-21-oic acid methyl esterin 25 ml. of anhydrous ether. The reaction mixture was heated underreflux for 1 hour. The excess lithium aluminum hydride was destroyed bythe addition of ethyl acetate. Water was added until a pasty mass oflithium salts were formed. The supernatant liquid was decanted andevaporated yielding a residue consisting essentially of the 3-pyrrolidylenamine of 6u,16a-dimethyl-115,21-dihydroxy-4,17(20)-pregnadien-3-one.

This residue was dissolved in 20 ml. of methanol containing 2 ml. of 5%aqueous sodium hydroxide. After 1.5 hours at 26 the solution wasneutralized with acetic acid, the methanol distilled and the residueextracted with methylene chloride and the extracts chromatographed overmagnesium silicate eluted with hexanes containing increasing proportionsof acetone. Hexanes plus 20% acetone eluted 6a,l6adimethyl-11/3,21-dihydroxy-4,l7 (20)-pregnadien-3-one.

PREPARATION 12 6a,] 6m-dime1thyl-J 1 3,21 -d1"hydroxy-] ,4 ,1 7(20pregnatrien-Z-one 2 gm. of corn steep liquor containing about 48% solidsand 1 gm. of glucose were added to tap water to make 100 ml. of totalliquid volume in a 250 ml. Erlenmeyer flask. The pH was adjusted to 4.8with sodium hydroxide solution and the resulting material sterilized for45 minutes at 15 pounds per square inch steam pressure. This medium wasinoculated with Septomyxa afiinis (A.T.C.C. 7637) and the inoculatedflask then placed on a rotary shaker where incubation was allowed toproceed for 2 days with the flask being rotated around a one-half inchradius at approximately 330 r.p.m. A substrate of 10 mg. of6a,16adimethyl-115,21-dihydroxy-4,

l7(20)-pregnadien-3-one dissolved in a minimum amount of propyleneglycol was introduced slowly with constant swirling of the flask tominimize precipitation of the steroid. 10 mg. of3-ketobisnor-4-cholen-22-al as the bioconversion assistant was dissolvedin 1 ml. of propylene glycol and added to the substrate-medium mixture.Bioconversion was then permitted to proceed for 4 days, the flask beingshaken throughout this period. At the end of this time, the steroidfraction was extracted with methylene chloride. The extracts were Washedtwice with sodium bicarbonate solution and twice with Water and thendried with sodium sulfate. The washed extract was freed of solvent andthe residue was triturated with ether. The residue was crystallized fromethylene dichloride to give substantially pure 60,16oc-diIIl6thYl-l113,21-dihydroxy- 1,4, 17 (20 -pregnatrien-3-one.

PREPARATION 13 6a,16a-dimethyl-1Ifl,21-dihydroxy-4,1 7(20) -pregnadien-3-0112 21-acetate A solution of mg. of60:,16ot-di1116thYI-1IKLZI-dihydroxy-4,17(20)-pregnadien-3-one in 1 ml.of pyridine and 1 ml. of acetic anhydride was maintained at roomtemperature for 17 hours. Crushed ice was added and the precipitate of6a,16a-dimethyl-11/3,21-dihydroxy- 4,17(20)-pregnadien-3 one 21-acetatewas extracted with 3 ten-milliliter portions of methylene chloride. Themethylene chloride extracts were dried over anhydrous sodium sulfate,evaporated and the residue recrystallized from a mixture of ethylacetate and hexanes to give 6:1 .,16a-diII16thYl-l1B,21-dihyd1OXy 4, 1720 -pregnadien- 3-one ZI-acetate.

Similarly, 60,160t dimethyl 11,6,21 dihydroxy 1,4,17(20)-pregnatrien-3-one is converted with acetic anhydride to6a,16a-dimethyl-11,8,21-dihydroxy-1,4,l7(20)- pregnatrien-3-one21-acetate.

PREPARATION 14 600,160L-dil716lh31l-1 10,1 70:,21-trihydr0xy-4-pregnene-3,20-di0ne 21 -acetate 100 mg. of 6ct,16cc dimethyl 115,21 dihydroxy-4,17(20)-pregnadien-3-one 21-acetate were dissolved in 6 ml. of tertiarybutyl alcohol and 0.05 ml. of pyridine. To this mixture was added 1.6mg. of osmium tetroxide and 0.8 mM. of N-methylmorpholine oxide peroxidein tertiary butyl alcohol. After stirring at 26 C. for 2 hours, excesssodium hydrosulfite solution was added. The solvent was distilled in avacuum and the product extracted with methylene dichloride. The materialwas chromatographed over magnesium sulfate (Florisil) and the fractionconsisting of hexanes (Skellysolve B) with 15% acetone was separated,and evaporated to give 611,16 dimethyl 11/3,17oz,21 trihydroxy 4pregnene- 3,20-dione 21-acetate.

Similarly, under exactly the same conditions, 60L,160C- dimethyl-118,21-dihydroxy-L4,17(20) pregnatrien-3-one 21-acetate was oxidativelyhydroxylated with osmium tetroxide and N-methylmorpholine oxide peroxideto 606,16OL-dlI1'1611hY1-11,8,170L,2 1-trihydroxy 1,4-pregnadiene-3,20-dione ZI-acetate.

PREPARATION 15 6a,16ot-dimethyl-I],8,1 704,21 -trihydr0xy-4-pregn ene-3,20-di0ne A solution was prepared containing 0.5 gm. of60:,16adimethyl-11;3,17a,21-trihydroxy 4 pregnene 3,20-dione 21-acetatein 25 ml. of methanol. This solution was purged with oxygen freenitrogen for a period of 5 minutes and then a similarly oxygen purgedsolution of 0.250 gm. of potassium bicarbonate dissolved in 1 ml. ofmethanol and 1 ml. of water was added. The mixture was maintained for 3hours in a nitrogen atmosphere, then neutralized with hydrochloric acid,poured into 200 ml. of ice water and the thus-obtained mixture extractedwith 9 4 fifty-milliliter portions of methylene chloride. The methylenechloride extracts were combined, washed several times with water, driedover anhydrous sodium sulfate and evaporated to give6a,16a-dimethyl-11fi,17u,21-trihydroxy-4-pregnene-3,20-dione.

Similarly, 60:,161! dimethyl 115,17a,21 trihydroxy-1,4-pregnadiene-3,ZO-dione Zl-acetate was hydrolyzed with potassiumbicarbonate to 6a,16a-dimethyl-11B,17a, 21trihydroxy-1,4-pregnadiene-3,20-dione.

PREPARATION 16 6a,16a-dimethyl-l 711,21 -dihydrxy-4,9(1 1 )-pregnadiene-3,20-di0ne 21 -acetare To a solution of 8.5 gm. of60,16ot-(11I116EhYl-1lB,17ot,21 trihydroxy-4-pregnene-3,ZO-dione21-acetate in 42.5 ml. of pyridine was added 5.63 gm. ofN-bromoacetamide. After standing at room temperature for a period of 15minutes, the reaction solution Was cooled to 5 to C. and sulfur dioxidegas was passed over the surface of the solution while shaking the flaskuntil the solution gave no color with acidified starch-iodide paper.During the addition of the sulfur diom'de, the reaction mixture becamewarm. The temperature was kept below 30 C. by external cooling and byvarying the rate of sulfur dioxide addition. Thereafter to the reactionmixture was added 400 ml. of ice Water and the resulting precipitatecollected by filtration. This material was recrystallized from a mixtureof acetone and hexanes (Skellysolve B) to give 611,160 dimethyl 17,21dihydroxy 4,9(11)-pregnadiene-3,20-dione 21-acetate.

Following the procedure of Preparation 16, but substituting6a,16a-dimethyl-1 1,8, 1711,21-trihydroxy-1,4-pregnadiene-3,20-dione21-acetate as the starting compound, there was thus produced6a,16a-dimethyl-17a,21-dihydroxy-1,4,9(11)-pregnatriene-3,20-dione2l-acetate.

PREPARATION 17 6a,]oat-dimethyl-9a-br0m0-1 1[3,17a,21-trihydr0xy-4-pregnene-3,20-di0ne 21-acemte To a solution of 5.68 gm. of6u,16x-dimethyl-17u,21-

' dihydroxy-4,9(1 1 -pregnadiene-3,20-dione 21 acetate in 100 ml. ofmethylene chloride and 250 ml. of tertiary butyl alcohol was added asolution of 14 ml. of 72% perchloric acid in 100 ml. of water followedby a solution of 2.34 gm. of N-bromoacetamide in 60 ml. of tertiarybutyl alcohol. After stirring the reaction mixture for 15 minutes, asolution of 2.8 gm. of sodium sulfite in 140 ml. of water was added andthe reaction mixture was concentrated to a volume of about 500 ml. underreduced pressure at about 50 C. The concentrate was cooled in an icebath and while stirring 500 ml. of water was added. After stirring for aperiod of 1 hour, the precipitated product was isolated by filtration,and the cake washed with water and air-dried to give6rx,16oz-din16thyl-9oz bromo l1 8,17a,2l trihydroxy 4pregnene-3,20-dione 21-acetate.

Following the procedure of Preparation 17, but substituting 6a,16adimethyl 170;,21 dihydroxy-1,4,9(11)- pregnatriene-3,20-dione 21-acetateas the starting compound, there Was thus produced6a,l6a-dimethyl-9abromo-l 1B,17a,21-trihydroxy-1,4pregnadiene-3,20-dione 21-acetate.

Substituting N-chlorosuccinimide for the N-bromoacetamide in thereactions described in Preparation 17 is productive of the corresponding9a-chloro compounds, i.e., 6d,].60t-d1mtl1yl-9Ct-Ch1OI'O-11fi,17a,21-trihydroxy-1,4- pregnadiene-3,20-dione 2l-acetate and6d,160t-dlm$thy1 9a-chloro-11B,17a,21-trihydroxy 4 pregnene-3,20-dione21-acetate.

PREPARATION 18 60c 16oz dimethyl 913,115 epoxy 1,7u,21dihydr0xy-4-pregnene-3,20-di0ne 21 -acelate To a solution of 6.78 gm. of6a,16a-dimethyl-9-bromo 11 ,9,17a,21 trihydroxy 4 pregnene 3,20 dione2l-acetate in 175 ml. of acetone was added 6.78 gm. of potassium acetateand the resulting suspension was heated under reflux for a period of 17hours. The mixture was then concentrated to appnoximately 60 ml. atreduced pressure on the steam bath, diluted with Water and extractedwith methylene chloride. The methylene chloride extracts were combined,washed with water, dried over anhydrous sodium sulfate and evaporated.The residue was redissolved in methylene chloride and chromatographedover 500 gm. of magnesium silicate (Florisil). The column was elutedwith l-liter portions of hexanes (Skellysolve B) containing increasingproportions of acetone. There was thus eluted 61x,16u-dimethyl 95,113epoxy 1711,21 dihydroxy 4 pregnene 3,21-dione 21-acetate which was freedof solvent by evaportaion of the eluates.

Following the procedure of Preparation 18, but substituting 6u,16 xdimethyl 9a bromo-11fi,17a-21-t1ihydroxy-1,4-pregnadiene-3,20-dione21-acetate as the starting compound, there was thus produced'6a,l6a-dimethyl- 95,11 8 epoxy 17a,21 dihydroxy 1,4 pregnadiene3,20-dione 21-acetate.

PREPARATION 19 604,16oc dimethyl c fluoro 11/8,17a,21 trihydroxy4-pregnene-3,20-dione 21 -acetate To approximately 1.3 gm. of hydrogenfluoride contained in a polyethylene bottle and maintained at minus 60C. was added 2.3 ml. of tetrahydrofuran and then a solution of 500 mg.of 6a,16adimethyl-9B,1IB-epoxy- 170,21-dihydroxy-4-pregnene-3,20-dione2l-acetate in 2 ml. of methylene chloride. The steroid solution wasrinsed in with an additional 1 ml. of methylene chloride. The light redcolored solution was then kept at approximately minus 30 C. for 1 hourand at minus 10 for 2 hours. At the end of this period it was mixedcautiously with an excess of cold sodium bicarbonate solution and theorganic material extracted with additional methylene chloride. Thecombined extracts werewashed with water, dried over anhydrous sodiumsulfate and concentrated to approximately 35 ml. The solution waschromatographed over gm. of magnesium silicate (Florisil). The columnwas developed with 260-m1 portions of hexanes (Skellysolve B) containingincreasing proportions of acetone. There was thus eluted 611,160:-dimethyl 9a fluoro 11,6,l7a,21 trihydroxy 4 pregnene-3,20-dioneZI-acetate which was freed of solvent by evaporation of the 'eluatefractions.

Following the procedure of Preparation 19, but substituting 6a.,160tdimethyl 913,115 epoxy 17a,21-dihydroxy-1,4-pregnadiene-3,20-dione21-acetate as the starting compound, there was thus produced6a,l6a-dimethyl- 9oz fluoro 1lfi,17oc,21 trihydroxy 1,4 pregnadiene3,20-dione 21-acetate.

PREPARATION 20 6a,]6a dimethyl 90c fluoro :,21 dihydroxy 4pregnene-3,11,20-trione 21 -acetate A solution of 1 ml. of acetic acid,50 mg. of 601,160:- dimethyl 9a fluoro llfi,l7a,2l trihydroxy 4pregnene-3,20-dione 21-acetate, 20 mg. of chromic anhydride and 1 drop(approximately 50 mg.) of water was shaken several times and thenmaintained at room temperature for 4 hours. Then, it was poured into 10ml. of water and refrigerated for 20 hours at about 5 C. The steroidwhich separated from the aqueous mixture was collected on filter paperand dried to give 60:, 16a dimethyl 9a fluoro 17a,21 dihydroxy 4pregnene-3,11,20trione 21-acetate.

Following the procedure of Preparation 20, but substituting 60;,16adimethyl 9o: fluoro 11[3,17a,21-trihy droxy-l,4-pregnadiene-3,ZO-dione21-acetate as the starting compound, there was thus produced6a,l6adimethyl 9a fluoro 17a,21 dihydroxy 1,4 pregnadiene 3,11,20-trione ZI-acetate.

PREPARATION 21 6a,16u dimethyl 90c fluoro 11fi,17a,21 trihydroxy4-pregnene-3,20-di0ne 3.25 gm. of 6a,l6a-dimethyl-9u-fluoro 11fi,17ot,21-trihydroxy-4-pregnene-3,ZO-dione 21-acetate' was dissolvedin 325 ml. of methanol, previously urged of air-oxygen by passingnitrogen through it for 10 minutes and thereto was added a solution of1.63 gm. of potassium bicarbonate in 30 ml. of water, similarly purgedof oxygen. The mixture was maintained at room temperature for 5 hours ina nitrogen atmosphere and then neutralized with 2.14 ml. of acetic acidin 40 ml. of water. The mixture was concentrated to approximatelyonethird volume at reduced pressure on a 60 C. water-bath, 250 ml. ofwater was then added and the mixture chilled. The crystalline productwas collected on a filter, washed with water and dried to give6ot,lGoa-dimthYl-9oz-flll0l0-111S,17,21-trihydroxy-4-pregnene-3,ZO-dione.

Following the procedure of Preparation 21, but substituting 60,16adimethyl 9a fluoro-1 lfi,17a,21-trihydroxy-l,4-pregnadiene-3,ZO-dione21-acetate as the starting compound, there was thus produced60.,16ct-diIfiGiLhYl- 9a fluoro 11,8,l7oc,21 trihydroxy 1,4pregnadiene-3,2()-dicne.

Similarly, 6a,l6cc dimethyl 9oz fluorol7a,21-dihydroxy-4-pregnene-3,11,20-trione 21-acetate is hydrolyzed to601,164 dimethyl-Sa-fluorm17a,Z1-dihydroxy-4-preg. nene-3,11,20-trioneand 6a,16a-dimethyl-9u-iiuoro-17a, 21 dihydroxy 1,4 pregnadiene 3,11,20trione ZI-acetate is hydrolyzed to 60,16cc dimethyl 9a-fiuoro- 170:,21dihydroxy 1,4 pregnadiene 3,11,20 trione. The corresponding 9ct-chlorocompounds are similarly prepared by hydrolysis of their 21-acetates,e.g., 606,160tdimethyl 9a chloro 1lfi,l7a,21 trihydroxy 1,4pregnadiene-3,20-dione and 60c,16ct-diIX16tl'1Yl-9u-Chl0f0- 11/3,17a,21trihydroxy 4 pregnene-3,20-dione are prepared from6a,liSOL-dlmMhYl-Qa-ChlOIO'I1B,l70t,21llflhydroxy-1,4-pregnadiene-3,20dioneZl-acetate and from 60:, 16a dimethyl 90c chloro 11B,l7ot,21 trihydroxy4-pregnene-3,20-dione 21-acetate, respectively.

EXAMPLE 1 60:,160; dimethyl 11fi,17ot,21 trihydroxy 4 pregnene-3,20-dine21 -m ethanesulf0nate A solution was prepared containing 1 gm. (2.6 mM.)of 60:,16m dimethyl-l1,8,17a,21-trihydroXy-4pregnene-3, 20-dione in 7ml. of pyridine. This solution was cooled to 0 C. and treated with 0.3ml. of methanesulfonyl chloride. The solution was maintained at O to C.for a period of 2 hours, after which it was diluted with water andextracted with 3 25-rnilliliter portions of methylene chloride. Theextracts were combined, washed with cold dilute hydrochloric acid untilthe aqueous layer had a pH of 2 to 3, then washed again with cold sodiumbicarbonate solution, water and finally dried over anhydrous sodiumsulfate. Evaporation of the methylene chloride extract at reducedpressure left a residue of 6a,

12 one 21-methanesulfonate, 60c,l6zx-dim6ihYl-9a-Chl0rO-llfl,170:,21-trihydroxy-4-pregnene-3,ZO-dione 21-methanesul fonate and6a,16a-dimethyl-9a-chloro-11B,17oc,21-trihydroxy-l,4-pregnadiene-3,20-dione21-methanesulfonate,

respectively.

EXAMPLE 2 6a,16cx-dimethyl-11,8,1 7u-dihydroxy-21-i0d0-4-pregnene-3,20-dione The crude 6a,16a-dimethyl-11 8,17a,21-trihydroxy-4-pregnene-3,20'-dione ZI-methanesulfonate described in Example l wasdissolved in 15 ml. of acetone and treated with a solution of 1 gm. ofsodium iodide in 10 ml. of acetone. The mixture was heated under refluxwith stirring for a period of 15 minutes, the heat then reduced and themixture concentrated to one-third volume at reduced pressure. Ice andwater were added and the precipitated product collected on a filter,washed with water and dried to yield6a,l6a-dimethyl-11B,17u-dihydroxy-21-iodo-4-pregnene-3,ZO-dione.

Similarly, 6tx-16et-dimethyl-1 1B, l7u,21-trihydroxy-1,4-pregnadiene-3,20-dione 21-methanesulfonate, 604,16a-dimethyl-9a-fluoroll/3,17oc,21 trihydroxy-1,4-pregnadicue-3,20-dione 21-methanesulfonate,60:,16oc-difll6thYl-9afluoro-11B,17 x,21-trihydroxy-4-pregnene 3,20dione 21- methanesulfonate, 6:1,160t-dl1116lhYl-96t-ChlOIO-l15,170L,21-trihydroxy-4-pregnene 3,20 dione ZI-methanesulfonate, and6a,l6a-dimethyl 9c: chloro-11,6,17a,21-trihydroxy-1,4-pregnadiene-3,ZO-dione 21-methanesulfonate are converted to6u,16ct-clirnethyl-11B,l7a-dihydroxy-2l-iodo-l,4-pregnadiene-3,20-dione, 6a,16a-dimethyl 9a fluoro-11B,17e-dihydroxy-21-iodo-1,4-pregnadiene 3,20 dione, 6a,160C-dlm6ll1yl-9d-fluOI'O-l 1fl,17a-dihydroxy 21 iodo-4-pregnene-3,20-dione, 60c, l6a-dimethyl-9a-chloro-1 113,170;-dihydroxy-21-iodo-4-pregnene-3,ZO-dione and 6a,l6a-dimethyl-9ot-chloro-115,l7a-dihydroxy-2l-iodo 1,4 pregnadiene3,2G-dione, respectively.

EXAMPLE 3 6 (1,1 6 ot-dimezhyl-l 1 5,1 7a-dihydr0xy-4-pregnene- 3 ,ZO-dione 150 mg. of6a,l6a-dimethyl-115,17a-dihydroxy-2liodo-4-pregnene-3,20-dione wasslurried with 5 m1. of acetic acid and stirred for a period of 45minutes. Then an aqueous solution of 256 mg. of sodium thiosulfatepentahydrate was added causing the iodine color to disappear. Additionalwater was added (50 ml.) and the mixture extracted with 3 25-milliliterportions of methylene chloride. The methylene chloride extracts werecombined, washed with water and cold sodium bicarbonate solution untilthe acetic acid was neutralized. After drying over anhydrous sodiumsulfate, the solution was concentrated to approximately 15 ml. andchromatographed over 10 gm. of magnesium silicate (Florisil). The columnwas developed with hexanes containing increasing proportions of acetone,to give substantially pure 6a,l6a-dimethyl-1lfi,l7a-dihydroxy 4pregame-3,20- dione.

Following the procedure of Example 3,60,16a-dimethyl-11fl,17a-dihydroxy-21-iodo 1,4 pregnadiene-3, 20-dionewas converted to 60,l6oc-dinl6thYl-l1,8,17a-dihydroxy-1,4-pregnadiene-3,ZO-dione.

Similary, 6a,16a-dimethyl cfluoro-llpLlh-dihyroxy-Z1-iodo-l,4-pregnadiene-3,ZO-dione,606,160t-dlll'16lhyl-9a-fluoro-11,8,17a-dihydroxy 21 iodo-4-pregnene-3,20-dione, 60c,16ot-dim6thyl-9a-ChlO1O-l 1B,17a-dihydroxy-21-iodo-4-pregnene-3,20-dione and6a,16ot-dimethyl-9uchloro-11/8,17u-dihydroxy-21-iodo 1,4 pregnadiene-3,20-dione were converted to 6a,16a-dimethyl-9a-fluoro-11,8,17a-dihydroxy-1,4-pregnadiene 3,20 dione,6a,16adimethyl-9u-fluoro-l1,8,17a-dihydroxy 4 pregnene-3,20- dione,60L,IGOL-(llll'lSlhYl-Qot-ChlOIO 11,8,17oc dihydroxy-4-pregnene-3,2G-dione and 611,1ot-ClllIlBthYl-9Dt-Chl0l'O-l1B,17oc-dihydroxy-1,4-pregnadiene-3,ZO-dione, respectively.

EXAMPLE 4 6 a,1 6a-dimethyl-1 7a-hydroxy-4-pregnene-3 ,1 1 ,20-

trione A mixture was prepared containing 0.3 gm. of 6a,16adimethyl-l1,8,17a-dihydroxy-4-pregnene-3,ZO-dione, 100 mg. of chromic anhyd-ride,10 ml. of glacial acetic acid and one-half ml. of water. This mixturewas stirred and thereupon maintained for 8 hours at room temperature.Thereafter the mixture was poured into 50 ml. of ice water, neutralizedby the addition of dilute sodium hydroxide and the thus-obtainedprecipitate collected on a filter and dried to give6a,l6a-dimethyl-17a-hydroxy-4- pregnene-3,11,20-trione.

Following the procedure of Example 4,6u,I6a-dimethyl-l113,17a-dihydroxy-1,4-pregnadiene-3,20-dione wasoxidized to 6a,l6a-dimethyl-17a-hydroxy-1,4-pregnadiene- 3,11,20-trione.Similarly, 6a,l6a-dimethyl 9afluorol1B,17a-dihydroxy-1,4-pregnadiene-3,ZO-dione,6a,16u-dimethyl-9u-fluoro-1lfi,l7u-dihydroxy 4 pregnene-3',20- dione,6a,16u-dimethyl-9u-chloro 11 3,17a dihydroxy-4- pregnene-3,20-dione and6a,16a-dimethyl-9a-ch1oro-11p,

14 17u-dihydroxy-1,4-pregnadiene-3,ZO-dione were converted to6a,16oc-dimethyl9a-fluoro-17a-hydroxy-1,4-pregnadiene-3,11,20-trione,6a,,16a-dimethy1 9a fluoro-17a-hydroxy-4-pregnene 3,11,20 trione,6oc,16a-dimethyl-9uchloro-l7u-hydroxy-4-pregnene-3,11,20-trione and60:,16ocdimethyl-9a-chloro-l7a-hydroxy 1,4 pregnadiene-3,11,

I 20-trione, respectively.

We claim: 6a,16a-dimethyl-9a-fluoro 115,170: dihydroxy-1,4-pregnadiene-3,20-dione.

References Cited in the file of this patent UNITED STATES PATENTS OTHERREFERENCES Arth et al.: J. Am. Chem. Soc., vol. 80, pages 3160-62 (June1958).

